CN107163149A - A kind of preparation method of lotus seed starch albumen composition - Google Patents
A kind of preparation method of lotus seed starch albumen composition Download PDFInfo
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- 229920002472 Starch Polymers 0.000 title claims abstract description 115
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- 238000006243 chemical reaction Methods 0.000 claims description 18
- CBMPTFJVXNIWHP-UHFFFAOYSA-L disodium;hydrogen phosphate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].OP([O-])([O-])=O.OC(=O)CC(O)(C(O)=O)CC(O)=O CBMPTFJVXNIWHP-UHFFFAOYSA-L 0.000 claims description 18
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- 229940024171 alpha-amylase Drugs 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
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- 239000002253 acid Substances 0.000 claims 3
- 238000007873 sieving Methods 0.000 claims 3
- 238000009413 insulation Methods 0.000 claims 2
- 238000001556 precipitation Methods 0.000 claims 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims 1
- 230000003139 buffering effect Effects 0.000 claims 1
- 238000001976 enzyme digestion Methods 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- 229910052698 phosphorus Inorganic materials 0.000 claims 1
- 239000011574 phosphorus Substances 0.000 claims 1
- 239000002131 composite material Substances 0.000 abstract description 26
- 150000003904 phospholipids Chemical class 0.000 abstract description 26
- 239000012467 final product Substances 0.000 abstract description 8
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 abstract description 6
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- 239000012460 protein solution Substances 0.000 description 2
- 150000003626 triacylglycerols Chemical class 0.000 description 2
- MPDGHEJMBKOTSU-YKLVYJNSSA-N 18beta-glycyrrhetic acid Chemical compound C([C@H]1C2=CC(=O)[C@H]34)[C@@](C)(C(O)=O)CC[C@]1(C)CC[C@@]2(C)[C@]4(C)CC[C@@H]1[C@]3(C)CC[C@H](O)C1(C)C MPDGHEJMBKOTSU-YKLVYJNSSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
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- 235000004213 low-fat Nutrition 0.000 description 1
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- QVLTXCYWHPZMCA-UHFFFAOYSA-N po4-po4 Chemical compound OP(O)(O)=O.OP(O)(O)=O QVLTXCYWHPZMCA-UHFFFAOYSA-N 0.000 description 1
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- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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Abstract
本发明提供了一种莲子淀粉‑蛋白复合物的制备方法，包括以下步骤：将莲子淀粉溶于酸性溶液中，加入淀粉酶进行酶解反应24～48h，接着调节体系为碱性，然后离心去除上清液得沉淀，将沉淀进行洗涤后去除上清液，干燥并粉碎得到莲子多孔淀粉；将磷脂与莲子多孔淀粉混合后后用微波‑超声波联合处理16～20min，得粗制的莲子多孔淀粉‑磷脂复合物；将蛋白、粗制的莲子多孔淀粉‑磷脂复合物及水混合后进行超高压处理，然后离心去除上清液得湿态的莲子淀粉‑蛋白复合物，接着将湿态的莲子淀粉‑蛋白复合物干燥至恒重后粉碎、过筛，得到终产品。本发明能制备出复合指数高的莲子淀粉‑茶多酚复合物。The invention provides a preparation method of lotus seed starch-protein complex, comprising the following steps: dissolving lotus seed starch in an acidic solution, adding amylase to carry out enzymatic hydrolysis reaction for 24-48 hours, then adjusting the system to be alkaline, and then centrifuging to remove The supernatant is precipitated, the precipitate is washed, the supernatant is removed, dried and crushed to obtain the lotus seed porous starch; the phospholipid and the lotus seed porous starch are mixed and then treated with microwave-ultrasonic wave for 16-20 minutes to obtain the crude lotus seed porous starch -Phospholipid complex; mix protein, crude lotus seed porous starch-phospholipid complex and water and perform ultra-high pressure treatment, then centrifuge to remove the supernatant to obtain wet lotus seed starch-protein complex, then wet lotus seed The starch-protein complex is dried to constant weight, crushed and sieved to obtain the final product. The invention can prepare lotus seed starch-tea polyphenol compound with high composite index.
Description
技术领域technical field
本发明涉及改性蛋白加工技术领域，特别涉及一种莲子淀粉-蛋白复合物的制备方法。The invention relates to the technical field of modified protein processing, in particular to a preparation method of lotus seed starch-protein complex.
背景技术Background technique
淀粉和蛋白质是两类重要的天然高分子聚合物，具有廉价易得、可降解和良好的生物相容性等特性。长期以来世界各国都十分重视淀粉和蛋白质资源的开发利用，尤其是二者的改性修饰一直是研究热点。在两者共存，一些物理化学条件适宜时，则发生共聚改性现象，即大分子上的部分基团可以互相连接复合，从而改善和赋予体系一些独特的功能性质、加工特性和及品质特性，最终扩大其在食品工业和其他工业中的应用，如可用作可食用膜、可用于开发低脂肪食品、药物缓释体系等。Starch and protein are two types of important natural polymers, which have the characteristics of cheap and easy to obtain, degradable and good biocompatibility. For a long time, all countries in the world have attached great importance to the development and utilization of starch and protein resources, especially the modification of the two has been a research hotspot. When the two coexist and some physical and chemical conditions are suitable, the phenomenon of copolymerization modification occurs, that is, some groups on the macromolecule can be connected to each other to compound, thereby improving and endowing the system with some unique functional properties, processing characteristics and quality characteristics. Finally, expand its application in the food industry and other industries, such as being used as an edible film, developing low-fat food, drug sustained release system, etc.
目前，淀粉-蛋白复合物的制备，主要有如下4种方法：At present, the preparation of starch-protein complexes mainly has the following four methods:
(1)干法反应通过控制自发美拉德反应来实现，将淀粉与蛋白粉以一定质量比混合后用去离子水溶解后冷冻干燥，冻干样品置于底部装有饱和KBr溶液，湿度为79％的反应器中，于60℃下反应数小时至几周；(1) The dry reaction is realized by controlling the spontaneous Maillard reaction. The starch and protein powder are mixed in a certain mass ratio and then dissolved in deionized water and then freeze-dried. The freeze-dried sample is placed at the bottom with a saturated KBr solution, and the humidity is 79% of the reactors were reacted at 60°C for several hours to several weeks;
(2)湿法反应淀粉与蛋白质的水溶液在一定条件下进行反应，蛋白质和淀粉按一定质量比混合，缓冲溶液稀释后置于密封离心管水浴加热接枝；(2) Wet method reaction starch and protein aqueous solution are reacted under certain conditions, and protein and starch are mixed by certain mass ratio, and buffer solution is placed in sealed centrifuge tube water bath heating grafting after dilution;
(3)电合成带有两根不锈钢电极的电池槽中装满质量分数淀粉和蛋白水溶液，通电后不时取下复合物凝胶，干燥即得复合物；(3) Electrosynthesis The battery tank with two stainless steel electrodes is filled with mass fraction starch and protein aqueous solution, and the complex gel is removed from time to time after electrification, and the complex is obtained after drying;
(4)挤压法淀粉与蛋白质按一定比例混合后采用螺杆机压机处理，并选择合适的挤压温度、水分含量、螺杆转速等相关技术参数。(4) Extrusion method Starch and protein are mixed in a certain proportion and processed by a screw press, and appropriate technical parameters such as extrusion temperature, moisture content, and screw speed are selected.
但是上述方法由于仅凭自发的美拉德反应，淀粉与蛋白不能充分复合，并且一般的制备法淀粉表面可与蛋白复合的颗粒成分(蛋白和脂质)较少，再加之条件要求严格，导致所制得的淀粉-蛋白复合物的复合率低，不便于用于复合物后期的使用与加工。But above-mentioned method owing to rely on spontaneous Maillard reaction only, starch and albumen can not be fully compounded, and the granule component (protein and lipid) that can be compounded with albumen on the surface of general preparation method starch is less, and condition requirement is strict in addition, causes The prepared starch-protein complex has a low complex rate, which is inconvenient for later use and processing of the complex.
发明内容Contents of the invention
本发明的目的在于克服了上述缺陷，提供一种莲子淀粉-蛋白复合物的制备方法，该方法制备出的莲子淀粉-蛋白复合物的复合率高。The object of the present invention is to overcome above-mentioned defect, provide a kind of preparation method of lotus seed starch-protein compound, the complex rate of the lotus seed starch-protein compound prepared by this method is high.
为了解决上述技术问题，本发明采用的技术方案为：In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:
本发明提供了一种莲子淀粉-蛋白复合物的制备方法，包括以下步骤：The invention provides a kind of preparation method of lotus seed starch-protein complex, comprises the following steps:
步骤1：将莲子淀粉溶于酸性溶液中，加入淀粉酶进行酶解反应24～48h，接着调节体系为碱性，然后离心去除上清液得沉淀，将沉淀进行洗涤后去除上清液，干燥并粉碎得到莲子多孔淀粉；Step 1: Dissolve lotus seed starch in an acidic solution, add amylase for enzymolysis reaction for 24-48 hours, then adjust the system to be alkaline, then centrifuge to remove the supernatant to obtain a precipitate, wash the precipitate, remove the supernatant, and dry And pulverize to obtain lotus seed porous starch;
步骤2：将磷脂与步骤1的莲子多孔淀粉混合后后用微波-超声波联合处理16～20min，得粗制的莲子多孔淀粉-磷脂复合物；Step 2: After mixing the phospholipid with the lotus seed porous starch in step 1, combined microwave-ultrasonic treatment for 16-20 minutes to obtain the crude lotus seed porous starch-phospholipid complex;
步骤3：将蛋白、步骤2所得粗制的莲子多孔淀粉-磷脂复合物及水混合后进行超高压处理，然后离心去除上清液得湿态的莲子淀粉-蛋白复合物，接着将湿态的莲子淀粉-蛋白复合物干燥至恒重后粉碎、过筛，得到终产品。Step 3: Mix the protein, the crude lotus seed porous starch-phospholipid complex obtained in step 2, and water, and then perform ultra-high pressure treatment, then centrifuge to remove the supernatant to obtain a wet lotus seed starch-protein complex, and then the wet state The lotus seed starch-protein complex is dried to constant weight, crushed and sieved to obtain the final product.
本发明还提供了一种莲子淀粉-蛋白复合物的制备方法，包括以下步骤：The present invention also provides a kind of preparation method of lotus seed starch-protein complex, comprises the following steps:
步骤1：将莲子淀粉溶于pH值为5.5的磷酸氢二钠-柠檬酸缓冲液中，于53～57℃条件下加热8～12min后加入α-淀粉酶与葡萄糖淀粉酶的复合酶，保温反应24～48h，接着利用1mol/L的氢氧化钠溶液调节体系为碱性，然后于5000r/min的转速下离心15min去除上清液得沉淀，将沉淀进行洗涤后去除上清液，将离心所得物于48～52℃常压条件下干燥至恒重然后粉碎、过筛得到莲子多孔淀粉，其中莲子淀粉与pH值为5.5的磷酸氢二钠-柠檬酸缓冲液的料液比为1∶1.5～2.5，料液比的单位为g/mL，所述1mol/L的氢氧化钠溶液与pH值为5.5的磷酸氢二钠-柠檬酸缓冲液的体积比为1∶10；Step 1: Dissolve lotus seed starch in disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5, heat at 53-57°C for 8-12 minutes, add the compound enzyme of α-amylase and glucoamylase, and keep warm React for 24 to 48 hours, then use 1mol/L sodium hydroxide solution to adjust the system to be alkaline, then centrifuge at a speed of 5000r/min for 15 minutes to remove the supernatant to obtain a precipitate, wash the precipitate, remove the supernatant, and centrifuge The resultant is dried to constant weight at 48-52°C under normal pressure, then crushed and sieved to obtain porous lotus seed starch, wherein the ratio of solid to lotus seed starch to disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5 is 1: 1.5～2.5, the unit of solid-liquid ratio is g/mL, the volume ratio of the sodium hydroxide solution of described 1mol/L and the disodium hydrogen phosphate-citric acid buffer solution that pH value is 5.5 are 1: 10;
步骤2：将磷脂与步骤1的莲子多孔淀粉混合后后用微波-超声波联合处理16～20min，得粗制的莲子多孔淀粉-磷脂复合物，用无水乙醇洗涤粗制的莲子多孔淀粉-磷脂复合物，然后冷冻干燥至恒重，得精制的莲子多孔淀粉-磷脂复合物，其中，所述莲子多孔淀粉与磷脂的质量比为1∶5～7，微波-超声波联合处理的条件包括微波功率为150～160W，超声波功率为350～400W，温度为70～90℃；Step 2: Mix the phospholipid with the lotus seed porous starch in step 1, and then use microwave-ultrasonic joint treatment for 16-20 minutes to obtain the crude lotus seed porous starch-phospholipid complex, wash the crude lotus seed porous starch-phospholipid with absolute ethanol The complex is then freeze-dried to constant weight to obtain a refined lotus seed porous starch-phospholipid complex, wherein the mass ratio of the lotus seed porous starch to phospholipid is 1:5-7, and the conditions for microwave-ultrasonic joint treatment include microwave power 150~160W, ultrasonic power 350~400W, temperature 70~90℃;
步骤3：将莲子蛋白与步骤2所得精制的莲子多孔淀粉-磷脂复合物分别溶解于蒸馏水后再混合得混合液，向混合液中添加pH为4.7的磷酸盐缓冲溶液，接着将添加有pH为4.7的磷酸盐缓冲溶液的混合液进行间歇式超高压处理20～30min，再进行洗涤，然后离心去除上清液得湿态的莲子淀粉-蛋白复合物，接着将湿态的莲子淀粉-蛋白复合物于48～52℃常压条件下干燥至恒重后粉碎、过筛，得到终产品，其中，所述莲子多孔淀粉-磷脂复合物与莲子蛋白的混合的质量比为1∶5～7，所述间歇式超高压处理的条件包括压力为200～250MPa，温度为60～80℃，每运行5min停歇30s。Step 3: dissolving the lotus seed protein and the refined lotus seed porous starch-phospholipid complex obtained in step 2 in distilled water and then mixing them to obtain a mixed solution, adding a phosphate buffer solution with a pH of 4.7 to the mixed solution, and then adding the mixed solution with a pH of The mixture of the phosphate buffer solution in 4.7 is subjected to intermittent ultra-high pressure treatment for 20-30 minutes, then washed, and then centrifuged to remove the supernatant to obtain the wet lotus seed starch-protein complex, and then the wet lotus seed starch-protein complex The product is dried to constant weight at 48-52°C under normal pressure, then pulverized and sieved to obtain the final product, wherein the mixed mass ratio of the lotus seed porous starch-phospholipid complex and lotus seed protein is 1:5-7, The conditions for the intermittent ultra-high pressure treatment include a pressure of 200-250 MPa, a temperature of 60-80° C., and a 30-s pause every 5 minutes of operation.
本发明的有益效果在于：(1)利用淀粉酶将普通莲子淀粉制备成莲子多孔淀粉来增大淀粉的表面积，利用微波-超声波联合处理辅助多孔淀粉吸附磷脂以增加淀粉表面的脂质成分，并利用非热力技术超高压提高蛋白质的表面疏水作用，大大增加淀粉与蛋白的复合指数，制备出高复合率的莲子淀粉-蛋白复合物；(2)本发明的制备方法未采用高温操作，可以有效保持淀粉与蛋白的结构，制备出的莲子淀粉-蛋白复合物的性能更佳，方便后续的使用与加工。The beneficial effects of the present invention are: (1) using amylase to prepare common lotus seed starch into lotus seed porous starch to increase the surface area of the starch, utilizing microwave-ultrasonic combined treatment to assist the porous starch to adsorb phospholipids to increase the lipid composition on the starch surface, and Utilize non-thermal technology ultra-high pressure to improve the surface hydrophobic effect of protein, greatly increase the composite index of starch and protein, and prepare the lotus seed starch-protein complex with high composite rate; (2) the preparation method of the present invention does not use high temperature operation, can effectively The structure of starch and protein is maintained, and the performance of the prepared lotus seed starch-protein complex is better, which is convenient for subsequent use and processing.
具体实施方式detailed description
为详细说明本发明的技术内容、构造特征、所实现目的及效果，以下结合实施方式详予说明。In order to describe in detail the technical content, structural features, achieved objectives and effects of the present invention, the following will be described in detail in conjunction with the embodiments.
本发明最关键的构思在于：通过将酶法制备的莲子多孔淀粉置于微波-超声波条件下来吸附磷脂以增大淀粉对蛋白质的吸附量，并利用超高压作用使淀粉与蛋白相互作用，进而制得复合率高的莲子淀粉-蛋白复合物。The most critical idea of the present invention is: the lotus seed porous starch prepared by the enzymatic method is placed under microwave-ultrasonic conditions to adsorb phospholipids to increase the adsorption amount of starch to protein, and use ultra-high pressure to make starch and protein interact, and then produce The lotus seed starch-protein compound with high compounding rate was obtained.
一种莲子淀粉-蛋白复合物的制备方法，包括以下步骤：A preparation method of lotus seed starch-protein complex, comprising the following steps:
步骤1：将莲子淀粉溶于酸性溶液中，加入淀粉酶进行酶解反应24～48h，接着调节体系为碱性，然后离心去除上清液得沉淀，将沉淀进行洗涤后去除上清液，干燥并粉碎得到莲子多孔淀粉；Step 1: Dissolve lotus seed starch in an acidic solution, add amylase for enzymolysis reaction for 24-48 hours, then adjust the system to be alkaline, then centrifuge to remove the supernatant to obtain a precipitate, wash the precipitate, remove the supernatant, and dry And pulverize to obtain lotus seed porous starch;
步骤2：将磷脂与步骤1的莲子多孔淀粉混合后后用微波-超声波联合处理16～20min，得粗制的莲子多孔淀粉-磷脂复合物；Step 2: After mixing the phospholipid with the lotus seed porous starch in step 1, combined microwave-ultrasonic treatment for 16-20 minutes to obtain the crude lotus seed porous starch-phospholipid complex;
步骤3：将蛋白、步骤2所得粗制的莲子多孔淀粉-磷脂复合物及水混合后进行超高压处理，然后离心去除上清液得湿态的莲子淀粉-蛋白复合物，接着将湿态的莲子淀粉-蛋白复合物干燥至恒重后粉碎、过筛，得到终产品。Step 3: Mix the protein, the crude lotus seed porous starch-phospholipid complex obtained in step 2, and water, and then perform ultra-high pressure treatment, then centrifuge to remove the supernatant to obtain a wet lotus seed starch-protein complex, and then the wet state The lotus seed starch-protein complex is dried to constant weight, crushed and sieved to obtain the final product.
本发明的工作原理及过程为：由于经申请人研究发现脂质有利于淀粉颗粒表面吸附更多的蛋白质，而极性的磷脂相比中性的糖脂、甘油三酸脂能吸附更多的蛋白质，故而，将莲子淀粉通过一定条件酶解制备成多孔淀粉，利用多孔淀粉表面积更大的优点来增加淀粉表面的脂质含量，进而增加淀粉对蛋白的吸附。微波-超声波联合处理辅助多孔淀粉吸附磷脂，使得纯磷脂通过渗透、扩散作用进入到多孔淀粉内部，增加淀粉表面的脂质成分，以利于淀粉表面吸附更多的蛋白质。超高压作为非热力技术在不破坏小分子物质的前提之下对蛋白质结构和性质产生影响，极大地提高蛋白质的表面疏水作用，结合淀粉得到高复合指数(高复合率)的复合物。The working principle and process of the present invention are as follows: due to the research by the applicant, it is found that lipids are conducive to the adsorption of more proteins on the surface of starch granules, and polar phospholipids can adsorb more proteins than neutral glycolipids and triglycerides. Therefore, the lotus seed starch is prepared into porous starch through enzymatic hydrolysis under certain conditions, and the advantage of larger surface area of porous starch is used to increase the lipid content on the starch surface, thereby increasing the adsorption of starch to protein. Microwave-ultrasonic combined treatment assists porous starch to adsorb phospholipids, so that pure phospholipids enter the interior of porous starch through penetration and diffusion, increasing the lipid composition on the starch surface, so as to facilitate the adsorption of more proteins on the starch surface. As a non-thermal technology, ultra-high pressure can affect the structure and properties of proteins without destroying small molecular substances, greatly improve the surface hydrophobicity of proteins, and combine with starch to obtain complexes with high recombination index (high recombination rate).
从上述描述可知，本发明的有益效果在于：(1)利用淀粉酶将普通莲子淀粉制备成莲子多孔淀粉来增大淀粉的表面积，利用微波-超声波联合处理辅助多孔淀粉吸附磷脂以增加淀粉表面的脂质成分，并利用非热力技术超高压提高蛋白质的表面疏水作用，大大增加淀粉与蛋白的复合指数，制备出高复合率的莲子淀粉-蛋白复合物；(2)本发明的制备方法未采用高温操作，可以有效保持淀粉与蛋白的结构，制备出的莲子淀粉-蛋白复合物的性能更佳，方便后续的使用与加工。As can be seen from the above description, the beneficial effects of the present invention are: (1) Utilize amylase to prepare common lotus seed starch into lotus seed porous starch to increase the surface area of starch, and utilize microwave-ultrasonic combined treatment to assist porous starch to adsorb phospholipids to increase the surface area of starch. Lipid components, and use non-thermal technology ultra-high pressure to improve the surface hydrophobic effect of protein, greatly increase the composite index of starch and protein, and prepare the lotus seed starch-protein complex with high composite rate; (2) the preparation method of the present invention does not use High temperature operation can effectively maintain the structure of starch and protein, and the performance of the prepared lotus seed starch-protein complex is better, which is convenient for subsequent use and processing.
进一步的，所述步骤1的酸性溶液为pH值为5.5的磷酸氢二钠-柠檬酸缓冲液，所述淀粉酶为α-淀粉酶与葡萄糖淀粉酶的复合酶，其中复合酶中α-淀粉酶的酶活为10 000U·g-1，葡萄糖淀粉酶的酶活为3 700U·g-1。Further, the acidic solution in step 1 is a disodium hydrogen phosphate-citrate buffer solution with a pH value of 5.5, and the amylase is a compound enzyme of α-amylase and glucoamylase, wherein α-amylase in the compound enzyme The enzyme activity of the enzyme is 10 000U·g -1 , and that of the glucoamylase is 3 700U·g -1 .
由上述描述可知，选择pH值为5.5的磷酸氢二钠-柠檬酸缓冲液作为酶解条件可以减小pH值的波动对酶解反应的影响，选择α-淀粉酶与葡萄糖淀粉酶的复合酶使得莲子淀粉被酶解得更充分。From the above description, it can be seen that choosing a disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5 as the enzymolysis condition can reduce the influence of pH fluctuations on the enzymolysis reaction, and choosing a compound enzyme of α-amylase and glucoamylase The lotus seed starch is enzymatically hydrolyzed more fully.
进一步的，所述步骤1的具体操作为：将莲子淀粉溶于pH值为5.5的磷酸氢二钠-柠檬酸缓冲液中，于53～57℃条件下加热8～12min后加入中温α-淀粉酶(10 000U·g-1)与葡萄糖淀粉酶(3 700U·g-1)的复合酶，保温反应24～48h，接着利用1mol/L的氢氧化钠溶液调节体系为碱性，然后离心去除上清液得沉淀，将沉淀进行洗涤后去除上清液，干燥至恒重然后粉碎、过筛得到莲子多孔淀粉，其中莲子淀粉与pH值为5.5的磷酸氢二钠-柠檬酸缓冲液的料液比为1∶1.5～2.5，料液比的单位为g/mL。Further, the specific operation of the step 1 is: dissolving lotus seed starch in disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5, heating at 53-57°C for 8-12 minutes, and then adding medium-temperature α-starch The compound enzyme of enzyme (10 000U·g -1 ) and glucoamylase (3 700U·g -1 ) was incubated for 24-48 hours, and then the system was adjusted to be alkaline with 1mol/L sodium hydroxide solution, and then removed by centrifugation. The supernatant is precipitated, the precipitate is washed, the supernatant is removed, dried to constant weight, then pulverized and sieved to obtain the lotus seed porous starch, wherein the lotus seed starch is mixed with disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5 The liquid ratio is 1:1.5-2.5, and the unit of the solid-liquid ratio is g/mL.
由上述描述可知，上述条件有助于酶解更充分且提高酶解效率，同时利用强碱溶液——氢氧化钠溶液调节体系pH以终止酶解反应，使用方便。It can be seen from the above description that the above conditions are conducive to more complete enzymatic hydrolysis and improved enzymatic hydrolysis efficiency. At the same time, the strong alkali solution-sodium hydroxide solution is used to adjust the pH of the system to terminate the enzymatic hydrolysis reaction, which is convenient to use.
进一步的，步骤2中，微波-超声波联合处理的条件包括：微波功率为150～160W，超声波功率为350～400W，温度为70～90℃。Further, in step 2, the conditions for the combined microwave-ultrasonic treatment include: microwave power of 150-160W, ultrasonic power of 350-400W, and temperature of 70-90°C.
由上述描述可知，将微波-超声波间歇处理的条件控制在上述范围有利于莲子多孔淀粉吸附更多磷脂且增加吸附的牢固性，进而便于后期吸附更多蛋白，进一步提高莲子淀粉-蛋白复合物的复合率。From the above description, it can be seen that controlling the conditions of microwave-ultrasonic intermittent treatment within the above range is conducive to the adsorption of more phospholipids by the lotus seed porous starch and increases the firmness of the adsorption, thereby facilitating the adsorption of more proteins in the later stage and further improving the lotus seed starch-protein complex. compound rate.
进一步的，在步骤2与步骤3之间还包括利用无水乙醇洗涤粗制的莲子多孔淀粉-磷脂复合物，然后冷冻干燥至恒重，得精制的莲子多孔淀粉-磷脂复合物。Further, between step 2 and step 3, the crude lotus seed porous starch-phospholipid complex is washed with absolute ethanol, and then freeze-dried to constant weight to obtain a refined lotus seed porous starch-phospholipid complex.
由上述描述可知，利用游离磷脂与莲子多孔-磷脂复合物在无水乙醇的溶解度的不同，将未游离磷脂洗去，避免游离磷脂直接与蛋白复合，以提高莲子淀粉-蛋白复合物的纯度。As can be seen from the above description, the difference in solubility between free phospholipids and lotus seed porous-phospholipid complexes in absolute ethanol is used to wash away free phospholipids, avoiding free phospholipids from directly complexing with proteins, and improving the purity of lotus seed starch-protein complexes.
进一步的，步骤3中，所述超高压处理为间歇式超高压处理，所述间歇式超高压处理的条件包括：压力为200～250MPa，温度为60～80℃，处理时间为20～30min，每运行5min停歇30s。Further, in step 3, the ultra-high pressure treatment is intermittent ultra-high pressure treatment, and the conditions for the intermittent ultra-high pressure treatment include: the pressure is 200-250 MPa, the temperature is 60-80 ° C, and the treatment time is 20-30 minutes, Stop for 30 seconds every 5 minutes.
其中，处理时间为每次运行时间之和。Among them, the processing time is the sum of each running time.
由上述描述可知，采用间歇式超高压处理及控制其处理条件在上述范围不仅可以提高蛋白的表面疏水作用增加后续复合率，还能起到杀菌效果。From the above description, it can be known that adopting intermittent ultra-high pressure treatment and controlling its treatment conditions within the above range can not only improve the surface hydrophobicity of the protein and increase the subsequent recombination rate, but also have a bactericidal effect.
进一步的，步骤3中，所述蛋白为莲子蛋白，所述莲子多孔淀粉-磷脂复合物与莲子蛋白的混合的质量比为1∶5～7。Further, in step 3, the protein is lotus seed protein, and the mass ratio of the lotus seed porous starch-phospholipid complex to lotus seed protein is 1:5-7.
由上述描述可知，采用莲子蛋白作为复合物的蛋白质，进一步提高莲子淀粉-蛋白复合物的复合率。From the above description, it can be seen that using lotus seed protein as the protein of the complex can further increase the compounding rate of the lotus seed starch-protein complex.
进一步的，步骤1及步骤3的干燥为于48～52℃常压条件下进行干燥。Further, the drying in step 1 and step 3 is carried out under normal pressure conditions of 48-52°C.
由上述描述可知，于48～52℃常压条件下进行干燥避免温度太高破坏淀粉及复合物的结构以保持复合物的性能的稳定性。It can be seen from the above description that drying is carried out under normal pressure conditions of 48-52° C. to avoid damage to the structure of the starch and the composite at a high temperature so as to maintain the stability of the performance of the composite.
本发明还提供一种莲子淀粉-蛋白复合物的制备方法，包括以下步骤：The present invention also provides a kind of preparation method of lotus seed starch-protein complex, comprises the following steps:
步骤1：将莲子淀粉溶于pH值为5.5的磷酸氢二钠-柠檬酸缓冲液中，于53～57℃条件下加热8～12min后加入中温α-淀粉酶(10 000U·g-1)与葡萄糖淀粉酶(3 700U·g-1)的复合酶，保温反应24～48h，接着利用1mol/L的氢氧化钠溶液调节体系为碱性，然后于5000r/min的转速下离心15min去除上清液得沉淀，将沉淀进行洗涤后去除上清液，将离心所得物于48～52℃常压条件下干燥至恒重然后粉碎、过筛得到莲子多孔淀粉，其中莲子淀粉与pH值为5.5的磷酸氢二钠-柠檬酸缓冲液的料液比为1∶1.5～2.5，料液比的单位为g/mL，所述1mol/L的氢氧化钠溶液与pH值为5.5的磷酸氢二钠-柠檬酸缓冲液的体积比为1∶10；Step 1: Dissolve lotus seed starch in disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5, heat at 53-57°C for 8-12 minutes, then add medium-temperature α-amylase (10 000U·g -1 ) Incubate and react with the compound enzyme of glucoamylase (3 700U·g -1 ) for 24-48 hours, then use 1mol/L sodium hydroxide solution to adjust the system to be alkaline, and then centrifuge at 5000r/min for 15min to remove the supernatant The supernatant liquid is precipitated, the precipitate is washed and the supernatant is removed, and the centrifuged product is dried to constant weight at 48-52°C under normal pressure, then crushed and sieved to obtain the lotus seed porous starch, wherein the lotus seed starch and the pH value are 5.5 The solid-liquid ratio of disodium hydrogen phosphate-citric acid buffer solution is 1:1.5～2.5, and the unit of solid-liquid ratio is g/mL, and the sodium hydroxide solution of described 1mol/L and the hydrogen phosphate dihydrogen phosphate that pH value is 5.5 The volume ratio of sodium-citrate buffer is 1:10;
步骤2：将磷脂与步骤1的莲子多孔淀粉混合后后用微波-超声波联合处理16～20min，得粗制的莲子多孔淀粉-磷脂复合物，用无水乙醇洗涤粗制的莲子多孔淀粉-磷脂复合物，然后冷冻干燥至恒重，得精制的莲子多孔淀粉-磷脂复合物，其中，所述莲子多孔淀粉与磷脂的质量比为1∶5～7，微波-超声波联合处理的条件包括微波功率为150～160W，超声波功率为350～400W，温度为70～90℃；Step 2: Mix the phospholipid with the lotus seed porous starch in step 1, and then use microwave-ultrasonic joint treatment for 16-20 minutes to obtain the crude lotus seed porous starch-phospholipid complex, wash the crude lotus seed porous starch-phospholipid with absolute ethanol The complex is then freeze-dried to constant weight to obtain a refined lotus seed porous starch-phospholipid complex, wherein the mass ratio of the lotus seed porous starch to phospholipid is 1:5-7, and the conditions for microwave-ultrasonic joint treatment include microwave power 150~160W, ultrasonic power 350~400W, temperature 70~90℃;
步骤3：将莲子蛋白与步骤2所得精制的莲子多孔淀粉-磷脂复合物分别溶解于蒸馏水后再混合得混合液，向混合液中添加pH为4.7的磷酸盐缓冲溶液，接着将添加有pH为4.7的磷酸盐缓冲溶液的混合液进行间歇式超高压处理20～30min，再进行洗涤，然后离心去除上清液得湿态的莲子淀粉-蛋白复合物，接着将湿态的莲子淀粉-蛋白复合物于48～52℃常压条件下干燥至恒重后粉碎、过筛，得到终产品，其中，所述莲子多孔淀粉-磷脂复合物与莲子蛋白的混合的质量比为1∶5～7，所述间歇式超高压处理的条件包括压力为200～250MPa，温度为60～80℃，每运行5min停歇30s。Step 3: dissolving the lotus seed protein and the refined lotus seed porous starch-phospholipid complex obtained in step 2 in distilled water and then mixing them to obtain a mixed solution, adding a phosphate buffer solution with a pH of 4.7 to the mixed solution, and then adding the mixed solution with a pH of The mixture of the phosphate buffer solution in 4.7 is subjected to intermittent ultra-high pressure treatment for 20-30 minutes, then washed, and then centrifuged to remove the supernatant to obtain the wet lotus seed starch-protein complex, and then the wet lotus seed starch-protein complex The product is dried to constant weight at 48-52°C under normal pressure, then pulverized and sieved to obtain the final product, wherein the mixed mass ratio of the lotus seed porous starch-phospholipid complex and lotus seed protein is 1:5-7, The conditions for the intermittent ultra-high pressure treatment include a pressure of 200-250 MPa, a temperature of 60-80° C., and a 30-s pause every 5 minutes of operation.
本发明的工作原理及过程为：由于经申请人研究发现脂质有利于淀粉颗粒表面吸附更多的蛋白质，而极性的磷脂相比中性的糖脂、甘油三酸脂能吸附更多的蛋白质，故而，将莲子淀粉通过一定条件酶解制备成多孔淀粉，利用多孔淀粉表面积更大的优点来增加淀粉表面的脂质含量，进而增加淀粉对蛋白的吸附。微波-超声波联合处理辅助多孔淀粉吸附磷脂，使得纯磷脂通过渗透、扩散作用进入到多孔淀粉内部，增加淀粉表面的脂质成分，以利于淀粉表面吸附更多的蛋白质。超高压作为非热力技术在不破坏小分子物质的前提之下对蛋白质结构和性质产生影响，极大地提高蛋白质的表面疏水作用，结合淀粉得到高复合指数(高复合率)的复合物。The working principle and process of the present invention are as follows: due to the research by the applicant, it is found that lipids are conducive to the adsorption of more proteins on the surface of starch granules, and polar phospholipids can adsorb more proteins than neutral glycolipids and triglycerides. Therefore, the lotus seed starch is prepared into porous starch through enzymatic hydrolysis under certain conditions, and the advantage of larger surface area of porous starch is used to increase the lipid content on the starch surface, thereby increasing the adsorption of starch to protein. Microwave-ultrasonic combined treatment assists porous starch to adsorb phospholipids, so that pure phospholipids enter the interior of porous starch through penetration and diffusion, increasing the lipid composition on the starch surface, so as to facilitate the adsorption of more proteins on the starch surface. As a non-thermal technology, ultra-high pressure can affect the structure and properties of proteins without destroying small molecular substances, greatly improve the surface hydrophobicity of proteins, and combine with starch to obtain complexes with high recombination index (high recombination rate).
从上述描述可知，本发明的有益效果在于：(1)利用淀粉酶将普通莲子淀粉制备成莲子多孔淀粉来增大淀粉的表面积，利用微波-超声波联合处理辅助多孔淀粉吸附磷脂以增加淀粉表面的脂质成分，并利用非热力技术超高压提高蛋白质的表面疏水作用，大大增加淀粉与蛋白的复合指数，制备出高复合率的莲子淀粉-蛋白复合物；(2)本发明的制备方法未采用高温操作，可以有效保持淀粉与蛋白的结构，制备出的莲子淀粉-蛋白复合物的性能更佳，方便后续的使用与加工。As can be seen from the above description, the beneficial effects of the present invention are: (1) Utilize amylase to prepare common lotus seed starch into lotus seed porous starch to increase the surface area of starch, and utilize microwave-ultrasonic combined treatment to assist porous starch to adsorb phospholipids to increase the surface area of starch. Lipid components, and use non-thermal technology ultra-high pressure to improve the surface hydrophobic effect of protein, greatly increase the composite index of starch and protein, and prepare the lotus seed starch-protein complex with high composite rate; (2) the preparation method of the present invention does not use High temperature operation can effectively maintain the structure of starch and protein, and the performance of the prepared lotus seed starch-protein complex is better, which is convenient for subsequent use and processing.
实施例1Example 1
1材料与方法1 Materials and methods
1.1材料1.1 Materials
莲子蛋白、莲子淀粉、磷酸氢二钠-柠檬酸缓冲液、中温α-淀粉酶(10 000U·g-1)与葡萄糖淀粉酶(3 700U·g-1)、l mol/L氢氧化钠、磷脂、无水乙醇、pH为4.7的磷酸盐缓冲溶液Lotus seed protein, lotus seed starch, disodium hydrogen phosphate-citrate buffer, medium temperature α-amylase (10 000U·g -1 ) and glucoamylase (3 700U·g -1 ), l mol/L sodium hydroxide, Phospholipids, ethanol, pH 4.7 phosphate buffer solution
1.2主要仪器1.2 Main instruments
DK-S24型电热恒温水浴锅，上海精宏实验设备有限公司；DK-S24 electric heating constant temperature water bath, Shanghai Jinghong Experimental Equipment Co., Ltd.;
TU-1901紫外-可见分光光度计，北京普析通用仪器有限责任公司；TU-1901 UV-Vis spectrophotometer, Beijing Puxi General Instrument Co., Ltd.;
5L-HPP-600MPa型超高压处理装置，包头科发高压科技有限责任公司；5L-HPP-600MPa ultra-high pressure processing device, Baotou Kefa High Pressure Technology Co., Ltd.;
SL-SM型微波超声波联合反应系统，南京顺流仪器制造有限公司；SL-SM Microwave Ultrasonic Combined Reaction System, Nanjing Shunliu Instrument Manufacturing Co., Ltd.;
TG16-WS型离心机，常州市金坛高科仪器厂；TG16-WS centrifuge, Changzhou Jintan Hi-Tech Instrument Factory;
BAS224S型电子分析天平，赛多利斯科学仪器(北京)有限公司。BAS224S electronic analytical balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.
1.3试验方法1.3 Test method
1.3.1一种莲子淀粉-蛋白复合物的制备方法，包括以下步骤：1.3.1 A preparation method of lotus seed starch-protein complex, comprising the following steps:
步骤1：称取25g莲子淀粉于于250mL三角瓶中，加入50mL pH值为5.5的磷酸氢二钠-柠檬酸缓冲液中，配制成淀粉乳悬液，于55℃条件下加热10min后加入中温α-淀粉酶(10000U·g-1)与葡萄糖淀粉酶(3 700U·g-1)的复合酶，保温反应24h，接着加入1mol/L的氢氧化钠溶液5mL以终止反应，然后于5000r/min的转速下离心15min去除上清液得沉淀，将沉淀进行洗涤后去除上清液，将离心所得物于50℃常压条件下干燥至恒重然后粉碎、过筛得到莲子多孔淀粉；Step 1: Weigh 25g of lotus seed starch into a 250mL Erlenmeyer flask, add 50mL of disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5 to prepare a starch emulsion suspension, heat at 55°C for 10min, then add to medium temperature The compound enzyme of α-amylase (10000U·g -1 ) and glucoamylase (3 700U·g -1 ) was incubated for 24h, then 5mL of 1mol/L sodium hydroxide solution was added to stop the reaction, and then heated at 5000r/L Centrifuge at a speed of 1 min for 15 min to remove the supernatant to obtain a precipitate, wash the precipitate and remove the supernatant, dry the centrifuged product at 50°C under normal pressure to a constant weight, then pulverize and sieve to obtain the lotus seed porous starch;
步骤2：将磷脂与步骤1的莲子多孔淀粉按5∶1的质量比混合后后用微波-超声波联合处理(微波-超声波联合处理的微波功率为150W，超声波功率为350W，温度为70℃)16min，得粗制的莲子多孔淀粉-磷脂复合物，用无水乙醇洗涤粗制的莲子多孔淀粉-磷脂复合物，然后冷冻干燥至恒重，得精制的莲子多孔淀粉-磷脂复合物；Step 2: After mixing the phospholipid and the lotus seed porous starch of step 1 in a mass ratio of 5:1, use microwave-ultrasonic joint treatment (the microwave power of the microwave-ultrasonic joint treatment is 150W, the ultrasonic power is 350W, and the temperature is 70°C) 16min to obtain the crude lotus seed porous starch-phospholipid complex, wash the crude lotus seed porous starch-phospholipid complex with absolute ethanol, then freeze-dry to constant weight to obtain the refined lotus seed porous starch-phospholipid complex;
步骤3：将莲子蛋白与步骤2所得精制的莲子多孔淀粉-磷脂复合物分别溶解于蒸馏水后再混合得混合液，向混合液中添加pH为4.7的磷酸盐缓冲溶液进行稀释，置于聚丙烯真空袋中用真空包装机封口，充分摇匀后放入进行间歇式超高压处理(间歇式超高压处理的压力为200MPa，温度为60℃，每运行5min停歇30s)20min，再进行洗涤，然后离心去除上清液得湿态的莲子淀粉-蛋白复合物，接着将湿态的莲子淀粉-蛋白复合物于50℃常压条件下干燥至恒重后粉碎、过筛，得到终产品，其中，所述莲子多孔淀粉-磷脂复合物与莲子蛋白的混合的质量比为1∶5。Step 3: The lotus seed protein and the refined lotus seed porous starch-phospholipid complex obtained in step 2 were dissolved in distilled water and then mixed to obtain a mixed solution, and a phosphate buffer solution with a pH of 4.7 was added to the mixed solution for dilution, and placed in polypropylene Seal the vacuum bag with a vacuum packaging machine, shake it well, put it into the intermittent ultra-high pressure treatment (the pressure of the intermittent ultra-high pressure treatment is 200MPa, the temperature is 60°C, and stop for 30s every 5 minutes) for 20 minutes, then wash, and then The supernatant was removed by centrifugation to obtain the wet lotus seed starch-protein complex, and then the wet lotus seed starch-protein complex was dried to a constant weight at 50°C under normal pressure, then crushed and sieved to obtain the final product, wherein, The mass ratio of the lotus seed porous starch-phospholipid complex to the lotus seed protein is 1:5.
1.3.2产品复合指数(复合率)的测定1.3.2 Determination of product composite index (composite rate)
通过测定离心后上清液中的莲子蛋白的量。用初始莲子蛋白的量与上清液中的莲子蛋白量的差值可计算出聚合物的复合率。上清液中莲子蛋白的浓度测定：By measuring the amount of lotus seed protein in the supernatant after centrifugation. The complex rate of the polymer can be calculated by using the difference between the amount of initial lotus seed protein and the amount of lotus seed protein in the supernatant. Determination of the concentration of lotus seed protein in the supernatant:
将超高压处理后的溶液离心得上清液，取1mL上清液用蒸馏水定容至100mL，以标准蛋白溶液做校正标准定量蛋白质，用分光光度计测278nm处吸收。The supernatant was obtained by centrifuging the ultra-high-pressure treated solution. Take 1 mL of the supernatant and dilute it to 100 mL with distilled water. Use the standard protein solution as a calibration standard to quantify the protein, and measure the absorption at 278 nm with a spectrophotometer.
根据下式计算莲子淀粉-蛋白复合物的复合指数：Calculate the complex index of lotus seed starch-protein complex according to the following formula:
CI＝(M initial-M supernatant)/M initial×100；CI=(M initial-M supernatant)/M initial×100;
式中:CI为莲子淀粉-蛋白复合物的复合指数；M initial为初始莲子蛋白的质量；M supernatant为上清液中莲子蛋白的质量。In the formula: CI is the complex index of lotus seed starch-protein complex; Minitial is the quality of initial lotus seed protein; M supernatant is the quality of lotus seed protein in the supernatant.
其中蛋白质含量M(mg)＝A×V×d/SLOPEstd×103；Among them, the protein content M (mg) = A × V × d/SLOPEstd × 103;
式中A为样品测试吸光度；V为样品提取体积；d为稀释因子，通常为1mL稀释成100mL，即稀释因子为100；SLOPEstd为蛋白溶液标准曲线的斜率。In the formula, A is the sample test absorbance; V is the sample extraction volume; d is the dilution factor, usually 1 mL is diluted to 100 mL, that is, the dilution factor is 100; SLOPEstd is the slope of the standard curve of the protein solution.
所测结果均重复测定三次，取均值，产品复合指数的测定结果见表1。The measured results were repeated three times, and the average value was taken. The results of the product composite index are shown in Table 1.
实施例2Example 2
其他同实施例1，不同之处在于莲子淀粉-蛋白复合物的制备，具体如下：Others are the same as Example 1, and the difference is the preparation of the lotus seed starch-protein complex, as follows:
步骤1：称取25g莲子淀粉于于250mL三角瓶中，加入50mL pH值为5.5的磷酸氢二钠-柠檬酸缓冲液中，配制成淀粉乳悬液，于55℃条件下加热10min后加入中温α-淀粉酶(10000U·g-1)与葡萄糖淀粉酶(3 700U·g-1)的复合酶，保温反应24h，接着加入1mol/L的氢氧化钠溶液5mL以终止反应，然后于5000r/min的转速下离心15min去除上清液得沉淀，将沉淀进行洗涤后去除上清液，将离心所得物于50℃常压条件下干燥至恒重然后粉碎、过筛得到莲子多孔淀粉；Step 1: Weigh 25g of lotus seed starch into a 250mL Erlenmeyer flask, add 50mL of disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5 to prepare a starch emulsion suspension, heat at 55°C for 10min, then add to medium temperature The compound enzyme of α-amylase (10000U·g -1 ) and glucoamylase (3 700U·g -1 ) was incubated for 24h, then 5mL of 1mol/L sodium hydroxide solution was added to stop the reaction, and then heated at 5000r/L Centrifuge at a speed of 1 min for 15 min to remove the supernatant to obtain a precipitate, wash the precipitate and remove the supernatant, dry the centrifuged product at 50°C under normal pressure to a constant weight, then pulverize and sieve to obtain the lotus seed porous starch;
步骤2：将磷脂与步骤1的莲子多孔淀粉按6∶1的质量比混合后后用微波-超声波联合处理(微波-超声波联合处理的微波功率为155W，超声波功率为375W，温度为80℃)18min，得粗制的莲子多孔淀粉-磷脂复合物，用无水乙醇洗涤粗制的莲子多孔淀粉-磷脂复合物，然后冷冻干燥至恒重，得精制的莲子多孔淀粉-磷脂复合物；Step 2: After mixing the phospholipid and the lotus seed porous starch of step 1 in a mass ratio of 6:1, use microwave-ultrasonic joint treatment (the microwave power of the microwave-ultrasonic joint treatment is 155W, the ultrasonic power is 375W, and the temperature is 80°C) 18min to obtain the crude lotus seed porous starch-phospholipid complex, wash the crude lotus seed porous starch-phospholipid complex with absolute ethanol, then freeze-dry to constant weight to obtain the refined lotus seed porous starch-phospholipid complex;
步骤3：将莲子蛋白与步骤2所得精制的莲子多孔淀粉-磷脂复合物分别溶解于蒸馏水后再混合得混合液，向混合液中添加pH为4.7的磷酸盐缓冲溶液进行稀释，置于聚丙烯真空袋中用真空包装机封口，充分摇匀后放入进行间歇式超高压处理(间歇式超高压处理的压力为225MPa，温度为70℃，每运行5min停歇30s)25min，再进行洗涤，然后离心去除上清液得湿态的莲子淀粉-蛋白复合物，接着将湿态的莲子淀粉-蛋白复合物于50℃常压条件下干燥至恒重后粉碎、过筛，得到终产品，其中，所述莲子多孔淀粉-磷脂复合物与莲子蛋白的混合的质量比为1∶6。Step 3: The lotus seed protein and the refined lotus seed porous starch-phospholipid complex obtained in step 2 were dissolved in distilled water and then mixed to obtain a mixed solution, and a phosphate buffer solution with a pH of 4.7 was added to the mixed solution for dilution, and placed in polypropylene Seal the vacuum bag with a vacuum packaging machine, shake it well, put it into the intermittent ultra-high pressure treatment (the pressure of the intermittent ultra-high pressure treatment is 225 MPa, the temperature is 70 ° C, every 5 minutes and stop for 30 seconds) for 25 minutes, then wash, and then The supernatant was removed by centrifugation to obtain the wet lotus seed starch-protein complex, and then the wet lotus seed starch-protein complex was dried to a constant weight at 50°C under normal pressure, then crushed and sieved to obtain the final product, wherein, The mass ratio of the lotus seed porous starch-phospholipid complex to the lotus seed protein is 1:6.
复合指数的评价方法同实施例1，产品复合指数的测定结果见表1。The evaluation method of composite index is the same as embodiment 1, and the measurement result of product composite index is shown in Table 1.
实施例3Example 3
步骤1：称取25g莲子淀粉于于250mL三角瓶中，加入50mL pH值为5.5的磷酸氢二钠-柠檬酸缓冲液中，配制成淀粉乳悬液，于55℃条件下加热10min后加入中温α-淀粉酶(10000U·g-1)与葡萄糖淀粉酶(3 700U·g-1)的复合酶，保温反应24h，接着加入1mol/L的氢氧化钠溶液5mL以终止反应，然后于5000r/min的转速下离心15min去除上清液得沉淀，将沉淀进行洗涤后去除上清液，将离心所得物于50℃常压条件下干燥至恒重然后粉碎、过筛得到莲子多孔淀粉；Step 1: Weigh 25g of lotus seed starch into a 250mL Erlenmeyer flask, add 50mL of disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5 to prepare a starch emulsion suspension, heat at 55°C for 10min, then add to medium temperature The compound enzyme of α-amylase (10000U·g -1 ) and glucoamylase (3 700U·g -1 ) was incubated for 24h, then 5mL of 1mol/L sodium hydroxide solution was added to stop the reaction, and then heated at 5000r/L Centrifuge at a speed of 1 min for 15 min to remove the supernatant to obtain a precipitate, wash the precipitate and remove the supernatant, dry the centrifuged product at 50°C under normal pressure to a constant weight, then pulverize and sieve to obtain the lotus seed porous starch;
步骤2：将磷脂与步骤1的莲子多孔淀粉按7∶1的质量比混合后后用微波-超声波联合处理(微波-超声波联合处理的微波功率为160W，超声波功率为400W，温度为90℃)20min，得粗制的莲子多孔淀粉-磷脂复合物，用无水乙醇洗涤粗制的莲子多孔淀粉-磷脂复合物，然后冷冻干燥至恒重，得精制的莲子多孔淀粉-磷脂复合物；Step 2: After mixing the phospholipid and the lotus seed porous starch of step 1 in a mass ratio of 7:1, use microwave-ultrasonic joint treatment (the microwave power of the microwave-ultrasonic joint treatment is 160W, the ultrasonic power is 400W, and the temperature is 90°C) 20min to obtain the crude lotus seed porous starch-phospholipid complex, wash the crude lotus seed porous starch-phospholipid complex with absolute ethanol, then freeze-dry to constant weight to obtain the refined lotus seed porous starch-phospholipid complex;
步骤3：将莲子蛋白与步骤2所得精制的莲子多孔淀粉-磷脂复合物分别溶解于蒸馏水后再混合得混合液，向混合液中添加pH为4.7的磷酸盐缓冲溶液进行稀释，置于聚丙烯真空袋中用真空包装机封口，充分摇匀后放入进行间歇式超高压处理(间歇式超高压处理的压力为250MPa，温度为80℃，每运行5min停歇30s)30min，再进行洗涤，然后离心去除上清液得湿态的莲子淀粉-蛋白复合物，接着将湿态的莲子淀粉-蛋白复合物于50℃常压条件下干燥至恒重后粉碎、过筛，得到终产品，其中，所述莲子多孔淀粉-磷脂复合物与莲子蛋白的混合的质量比为1∶7。Step 3: The lotus seed protein and the refined lotus seed porous starch-phospholipid complex obtained in step 2 were dissolved in distilled water and then mixed to obtain a mixed solution, and a phosphate buffer solution with a pH of 4.7 was added to the mixed solution for dilution, and placed in polypropylene Seal the vacuum bag with a vacuum packaging machine, shake it well, put it into the intermittent ultra-high pressure treatment (the pressure of the intermittent ultra-high pressure treatment is 250MPa, the temperature is 80°C, and stop for 30s every 5min) for 30min, then wash, and then The supernatant was removed by centrifugation to obtain the wet lotus seed starch-protein complex, and then the wet lotus seed starch-protein complex was dried to a constant weight at 50°C under normal pressure, then crushed and sieved to obtain the final product, wherein, The mass ratio of the lotus seed porous starch-phospholipid complex to the lotus seed protein is 1:7.
复合指数的评价方法同实施例1，产品复合指数的测定结果见表1。The evaluation method of composite index is the same as embodiment 1, and the measurement result of product composite index is shown in Table 1.
对比例comparative example
淀粉-蛋白复合物的干法反应制备：Dry reaction preparation of starch-protein complexes:
通过控制自发美拉德反应来实现。将淀粉与蛋白粉按1:7混合后用去离子水溶解调至6％(m/V))后冷冻干燥。冻干样品置于底部装有饱和KBr溶液，湿度为79％的反应器中，干热反应控制在60℃进行，反应持续5d。This is achieved by controlling the spontaneous Maillard reaction. Starch and protein powder were mixed at a ratio of 1:7, dissolved in deionized water and adjusted to 6% (m/V)), and then freeze-dried. The lyophilized samples were placed in a reactor with a saturated KBr solution at the bottom and a humidity of 79%. The dry heat reaction was carried out at 60°C and the reaction lasted for 5 days.
复合指数的评价方法同实施例1，产品复合指数的测定结果见表1。The evaluation method of composite index is the same as embodiment 1, and the measurement result of product composite index is shown in Table 1.
表1莲子淀粉-蛋白复合物复合指数(复合率)的测定结果Table 1 Determination results of lotus seed starch-protein complex composite index (composite rate)
由表1可知，本发明制备的莲子淀粉-蛋白复合物相比于对比例，其复合指数提高了1倍。It can be seen from Table 1 that the composite index of the lotus seed starch-protein composite prepared by the present invention is doubled compared with that of the comparative example.
综上所述，本发明提供的莲子淀粉-蛋白复合物的制备方法，利用淀粉酶将普通莲子淀粉制备成莲子多孔淀粉来增大淀粉的表面积，利用微波-超声波联合处理辅助多孔淀粉吸附磷脂以增加淀粉表面的脂质成分，并利用非热力技术超高压提高蛋白质的表面疏水作用，大大增加淀粉与蛋白的复合指数，制备出高复合率的莲子淀粉-蛋白复合物；本发明的制备方法未采用高温操作，可以有效保持淀粉与蛋白的结构，制备出的莲子淀粉-蛋白复合物的性能更佳，方便后续的使用与加工。In summary, the preparation method of lotus seed starch-protein complex provided by the present invention uses amylase to prepare ordinary lotus seed starch into lotus seed porous starch to increase the surface area of the starch, and uses microwave-ultrasonic combined treatment to assist the porous starch to absorb phospholipids to Increase the lipid composition on the starch surface, and use non-thermal technology ultra-high pressure to improve the surface hydrophobic effect of protein, greatly increase the composite index of starch and protein, and prepare the lotus seed starch-protein composite with high composite rate; the preparation method of the present invention does not The high temperature operation can effectively maintain the structure of starch and protein, and the prepared lotus seed starch-protein complex has better performance and is convenient for subsequent use and processing.
以上所述仅为本发明的实施例，并非因此限制本发明的专利范围，凡是利用本发明说明书所作的等效结构或等效流程变换，或直接或间接运用在其他相关的技术领域，均同理包括在本发明的专利保护范围内。The above is only an embodiment of the present invention, and does not limit the patent scope of the present invention. Any equivalent structure or equivalent process transformation made by the description of the present invention, or directly or indirectly used in other related technical fields, shall be the same as The theory is included in the patent protection scope of the present invention.
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